Analysis of Pesticides using HPLC

Muhammet Mammetkuliyev November 27, 2013

Abstract A high performance liquid chromatography (HPLC) method was developed to quantify the p,p -dichlorodiphenyltrichloroethane (DDT) dissolved in acetonitrile. Colorimetric detection was used with 236.4 nm incident light. The absorbance showed a linear increase in the concentration range of 10-25 ppm. A test solution with known concentration was analyzed with the developed method. The measured and actual concentrations of test solution diered by 4.91 ppm and the percent deviation is 37.3 %. The reason for this deviation is proposed.

Introduction
Dichlorodiphenyltrichloroethane (DDT) was rst synthesized in 1874 by a German chemistry student Othmar Zeidler. He was unaware of the insecticidal property of DDT. The available insecticides at the time were either expensive natural products or ineective insecticides. The only available insecticide was arsenic which is toxic to both animals and humans [1]. The 1

discovery of the insecticidal property of DDT was done in 1939 by Paul Hermann M uller. His rst experiment was on a y in a cage. Later, larger scale tests of the DDT against Colorado potato beetle, mosquito, louse and ea proved the eectiveness the insecticide [1]. DDT was patented in Switzerland, US, Australia and UK within 4 years of its discovery and mass production had begun. Its benets came with a cost, however. It was banned in Sweden in 1970 due to ecological reasons [2]. Its production slowed down such that only three dedicated factories are operational in the world and they are in US, France and India [2]. The main route for DDT intake in human is through ingestion. It has been proposed that the main parts in the body targeted by DDT is the nervous system and the liver [2]. In the liver, DDT is linked to the carcinogenic activity [2]. Another study elucidated the eect of DDT on the sperm health in men [3]. The key ndings from the study are that the decrease in sperm concentration and the decrease in the motility of the sperm in men are associated with the DDT presence in the body [3]. Gas chromatography and colorimetric methods are widely used for quantication of DDT. With gas chromatographic method, the electron capture detectors are good for quantication of DDT as they have electronegative chlorine atoms. Colorimetric methods, on the other hand, relies on the broad absorption of DDT in the range of 200-285 nm. In this study, high performance liquid chromatography (HPLC) used to identify and quantify the DDT dissolved in acetonitrile.

Experimental
The ideal wavelength of 236.4 nm to be used in HPLC was found using UV/Vis spectrometer. DDT was dissolved in acetonitrile and the absorption spectrum in 220-450 nm range was recorded. The absorbance due to acetonitrile was subtracted from the DDT solution. The HPLC method also used acetonitrile a mobile phase. Four standard solutions and the test solution were prepared using pure p,p - dichlorodiphenyltrichloroethane (Chem Service, Lot 63-45A, Purity 97.0%) with the concentration in the range of 10-25 ppm and 13.16 ppm, respectively. The ow rate for the HPLC runs were tested to get good peak separation in relatively short scan length. With the resultant solvent programming, the DDT peaks were eluted at 2.13 min.

Results and Discussion

The DDT peak areas for each standard solution concentrations are shown in Table 1. The linear t and the experimental data agree very well as shown in Figure 1. Our unknown solution peak area was 8385 and the measured concentration is thus 8.25 ppm compared with the actual value of 13.16 ppm. The dierence between the measured and actual concentrations is 4.91 ppm and the percent deviation is 37.3 %.

Concentration (ppm) Peak area

9.40

16.45

18.80

22.56

9361 15778 17893 21089

Table 1: DDT concentrations and peak areas at 236.4 nm.

There are two possible explanation for the observed disparity: either the test solution concentration is indeed 13.16 ppm and that the instrumental factors aected the measurements or that the test solution concentration is 8.25 ppm and a random error was done during the dilution. The former reason is not convincing since the standard curve shown in Figure 1 ts very well to the experimental data thus suggesting that the reproducibility of the tests with our instrument is very good. Moreover, analyzing the peak shapes of the standard and test solutions support this initial conclusion. All peaks have a Gaussian shape and are free from irregular features such as shoulders as shown in Figure 2. Therefore, a random error during the unknown preparation is thought to be the real cause of the disparity.

20000

Experimental data Linear fit. 2 y=894.0x+1008.3 (R = 0.9998)

Peak area (AU)

15000

10000

10

20 15 DDT concentration (ppm)

Figure 1: Absorbance of Cr(VI) standard solutions at 540 nm.

Conclusion
In this study, a method was developed to quantitatively analyze the pure DDT concentration. For the separation of DDT and its analysis, HPLC was used with acetonitrile as the mobile phase. The detector was set to 236.4 nm since DDT and acetonitrile absorptions are strong and negligible, respectively, at this wavelength. The linear t to the experimental DDT absorbance values vs. concentration were in very good agreement. Therefore, the 236.4 nm absorption of DDT molecules in the concentration range of 1025 ppm with acetonitrile as a solvent shows a linear behavior.

References
[1] World of Anatomy and Physiology. Gale Cengage, 2002. [2] DDT and its Derivatives. Technical report, World Health Organization, 1979. [3] S.E. Martenies and M.J. Perry. Environmental and occupational pesticide exposure and human sperm parameters: A systematic review. Toxicology, 307:66, 2013.

9.4 ppm std

16.45 ppm std

18.80 ppm std

22.56 ppm std

13.16 ppm test

Figure 2: DDT absorbance peaks for standard and test solutions at 236.4 nm.